• Restorative Dentistry and Endodontics
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• v.20 no.1
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• pp.71-95
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• 1995

Bacteria have been regarded as a one of major etiologic factors in root canal infections. In endodontic treatment the effective removal of pathogenic microorganisms in the root canal is the key to successful outcome. Bacterial cell wall components may play an important role in the development of pulpal and periapical disease. The purpose of this study was to evaluate the effect of sonic extracts of Streptococcus spp. on cultured L929 cells and to characterize cell wall protein profiles of Streptococcus spp. Streptococcus spp. were isolated from infected root canals and identified with Vitek Systems(Biomeriux, USA). Five streptococci, namely S. sanguis, S. mitis, S uberis, S. mutans (ATCC 10449) and S. faecalis (ATCC 19433) weere enriched in brain heart infusion broth. Cell pellets were sonicated and cell wall extracts were dialyzed and membrane filtered. Prepared cell wall proteins were applied to cultured L929 cell. The cell reaction were evaluated by monitoring DNA synthesis, cell numbers and the change of cell morphology. The total cell wall protein profiles of microorganisms were characterized by sodium dodecyl sulfate polyacrylamide-gel eledruphoresis(SDS-PAGE). DNA synthesis of L929 cells were reduced by the increasing concentration of sonic extracts. DNA synthesis was significantly suppressed in more than $50{\mu}g$/ml of sonic extract conentration in five streptococci. S. nutans (ATCC 10449) showed stronger suppression on DNA synthesis than remaining four streptococci, which had the similar effect on DNA synthesis. Analysis of DNA synthesis measured by [$^3H$]-thymidine uptake was more sensitvie method than cell counting. Sonic extracts affected the microscopic findings of L929 cells. The protein profiles indicated that all five strains shared two major proteins with molecular masses of 70.8 and 57.5 kD respectively. S. uberis and S. mutans shared common minor proteins of which molecular weights were 147.9 and 112.2 kD respectively. However some minor proteins were unique for S. mitis, S. uberis and S. faecalis.

• Restorative Dentistry and Endodontics
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• v.30 no.2
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• pp.138-144
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• 2005

This in vitro study monitored MMP-8 production on PMN by stimulated with the following three groups; Sonicated extracts of E. faecalis (SEF), SEF treated with $Ca(OH)_2$ (12.5mg/ml) for 7 days, and lipopolysac-charides(LPS) of E. coli. The level of MMP-8 in each group was immediately measured by ELISA. The data were analyzed with Kruskal-Wallis test and Mann-Whitney U test. In the SEF group, the level of production of MMP-8 was higher than the negative control group in low concentration ($0.05{\mu}g/ml$) of SEF (p < 0.05). but it decreased with an increase in the concentration of SEF (p < 0.05). In the case of SEF treated with $Ca(OH)_2$, all of the MMP levels were higher than negative control group (p < 0.05), but no statistical difference was found among the different SEF concentrations (p > 0.05). All of the levels in E. coli LPS were incresed with increasing concentrations (p < 0.05). According to this study we could summarized as follows: 1. MMP-8 was expressed at low level in untreated PMN group the levels of MMP-8 were upregulated in PMN stimulated by E. coli LPS groups. 2. In the SEF groups, the level of production of MMP-8 decreased with an increase in concentration of SEF (p < 0.05). So E. faecalis may have suppressive effect on the production of MMP-8 by PMN. 3. In the case of SEF treated with $Ca(OH)_2$, all of the MMP levels at different SEF concentrations were higher than untreated PMN group (p < 0.05), but no statistical difference was found among the different SEF concentrations (p > 0.05).

• Restorative Dentistry and Endodontics
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• v.30 no.1
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• pp.1-6
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• 2005

In order to examine the immunoresponse of host cells to Enterococcus faecalis, this in vitro study monitored the production of Interleukin-2 (IL-2), Interleukin-4 (IL-4) and Transforming growth factor-$\beta1\;(TGF-\beta1)$ in human lymphocytes. Lymphocytes were activated with PHA in the presence or abscence of sonicated extracts of E. Faecalis (SEF) and further incubated for 72 hours. The level of each cytokine was measured by ELISA. Data were analyzed with Kruskal-Wallis test and Mann-Whitney U test (P < 0.05). PHA-activated group did exhibit higher level of IL-2 and IL-4 than untreated control group. The levels of expression of both cytokines were significantly decreased following the treatment of high (25 ${\mu}g/ml$) and medium concentration (12.5 ${\mu}g/ml$)) of SEF (P > 0.05) than those of PHA activated group. But low concentration (5 ${\mu}g/ml$)) of SEF showed th similar level of IL-2 and IL-4 production as those of PHA activated group. $TGF-\beta1$ was unaffected by SEF treatment. These results suggested that E. faecalis may suppress IL-2 and IL-4 production by lymphocytes and this could be one of possible factors why E. faecalis are found frequently in the teeth with failed endodontic treatment.

• Natural Product Sciences
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• v.5 no.4
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• pp.181-185
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• 1999

Higenamine is a cardiotonic constituent of Aconite root, one of the most important oriental traditional medicine. Since Aconite root contains toxic aconitine alkaloids, variously processed roots have been often used. Much works have been done with the chemical significances concerning with the toxic aconitine alkaloids during the processing periods. However, effects of processing on higenamine have not yet been previously studied. In this paper, the extract pattern and the amounts of higenamine extracted with water from unprocessed and processed Aconite roots were compared. R-(+)-isomer was the only higenamine enantiomer detected although racemic higenamine was reported to be separated from Aconitum spp. Sonication for 1 hour resulted in higher higenamine extraction $(12.3\;{\mu}g/g)$ than boiling water extraction for 3 hours $(6.7\;{\mu}g/g)$ of unprocessed Aconite root. Extraction of not only higenamine but also most of the other components of unprocessed Aconite roots were reduced with boiling in water. Similarly, reduced extraction was observed with extracts of all three processed Aconite roots (Kyung-Po-Aconite root, Dang-Po-Aconite root and Huk-Peon-Aconite root) by either sonicated extraction or boiling water extraction.

• Journal of Periodontal and Implant Science
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• v.33 no.4
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• pp.585-597
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• 2003

Porphyromonas gingivalis has been implicated as an important periodontophathic bacterium in the etiology and progression of periodontal diseases. It has been reported that P.gingivalis may mediate periodontal destruction not only directly through its virulence factors, but also indirectly by including complex host mediated inflammatory reponses. The purpose of this study was t o evaluate the effects of P.gingivalis on the bone formation and resorption by osteoblasts. For this purpose, after determining the concentration below which sonicated P.gingivalis extracts (SPEs) have no cytotoxicity on mouse calvarial primary osteoblastic (POB) cells, we investigated the effects of SPEs on the alkaline phosphatase (ALP) activity, matrix metalloproteinase (MMP) expression (MMP-2, -9, 13), and prostaglandin $E_2$ ($PGE_2$) release in POB cells by treatment with SPEs below that concentration. The results were as follows; 1. SPEs showed no cytotoxic effect on POB cells up to a concentration of 1 ${\mu}m$/ml. 2. The treatment with SPEs reduced ALP activity in a dose-dependent manner in POB cells, In addition, when we investigated the effect of SPEs (1 ${\mu}m$/ml) on ALP activity for different exposure periods, statistically significant inhibition of ALP activity was shown at 2 days of exposure, and further significant inhibition occurred by extending the periods of exposure. 3. The treatment with SPEs stimulated the gene expression of MMP-9 in POB cells. 4. The pre-treatment with SPEs increased the amount of $PGE_2$ released in POB cells. In summary, the present study shows that P.gingivalis could inhibit osteogenesis and stimulate bone resorption not only by reducing ALP activity but also by increasing MMP-9 mRNA expression in osteoblasts, possibly through an endogenous $PGE_2$ pathway. In addition, our results suggest that if P.gingivalis affects osteoblasts in early differentiation stage, such effects by P. gingivalis could be irreversible.

• Journal of Naturopathy
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• v.12 no.1
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• pp.7-15
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• 2023

Background: As a result of analyzing the components of wild Conyza canadensis, it contains physiologically active ingredients, so it is necessary to identify the compound. Purposes: It was to study the compound's molecular structure; a previous study showed that C. canadensis contains antioxidant substances. Methods: The ultrasonic pulverized lysate of C. canadensis stem and leaves was first extracted with 90% methanol and then five organic solvents. Next, the extracts was fractionated by HPLC, LC/MS chromatography, and NMR analyzers identified the molecular structure. Results: 100 g of dry C. canadensis was sonicated in 90% methanol and concentrated under reduced pressure to 11.96 g of a crude extract. Then, this crude was extracted with five types of solvents to obtain 123.8 mg of n-hexane, 448.2 mg of dichloromethane, 1047.7 mg of ethyl acetate (EA), 2563.8 mg of butanol, and 7.04 g of water. The EA extracts were fractionated by LC-MS and then re-fractionated to obtain F1 to F20. Next, the F15 was further fractionated to obtain nine fine fractions. Finally, the F17 fraction was re-fractionated to obtain ten fine fractions. As a result of LC-MS and NMR spectrometer analysis of the F15-7, the structure of this compound was confirmed as 3,5-dicaffeoylquinic acid. As a result of examining the structures of the F17-4 and F17-5 fractions, Quercetin-3-o-β-galactose was identified. In addition, the form of the F17-10 was confirmed to be 1,3,4-tri-caffeoylquinic acid. Conclusions: This study demonstrated that C. canadensis contained phenolic antioxidants, and its utilization may be expected.

• Journal of Korean Ophthalmic Optics Society
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• v.15 no.4
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• pp.347-353
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• 2010

Purpose: An in vitro study was conducted to compare the lipid cleaning efficacy of care solutions on balafilcon A silicone hydrogel (SiHy) lens. Methods: Lipid spoilation was performed by incubating balafilcon A SiHy lenses in phosphate buffered saline (PBS) containing oleic acid, oleic acid methyl ester and cholesterol. Spoiled contact lenses rinsed with PBS were cleaned with surfactant cleaner, alcohol containing cleaner and multipurpose solution (MPS) respectively and repetitive spoilation and cleaning was conducted up to 14 times. To observe the cleaning effect of ultrasonic wave on the lipid deposit, each spoiled lens was ultrasonicated and then compared with non-sonicated lens. Lipids deposit on the contact lenses was extracted by methanol:chloroform (1:1, v/v) solution. High performance liquid chromatography was used to analyze and quantify lipid deposit extracts. Results: The effectiveness of alcohol containing surfactant cleaner on the lipid deposits was better than that of surfactant cleaner and MPS, and the cleaning efficacy was significantly higher in the ultrasonic wave treated group. Lipid deposits were not removed completely by contact lens care solutions so that lipid deposits increased continuously and cumulatively. Conclusions: The cleaning efficacy of contact lens care solutions was not satisfactory to remove lipid deposits on the SiHy lens that new cleaning products specially designed for SiHy lenses are needed to develop.

• Journal of Periodontal and Implant Science
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• v.29 no.4
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• pp.979-995
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• 1999

This study was investigated to observe the effect of Treponema denticola cell sonicates(TDC) and Treponema lecithinolyticum cell sonicates(TLC) on cytokine secretion and matix metalloproteinase-2(MMP-2) activation of cultured human gingival fibroblast. Several experiments were performed including $IL-1{\beta}$, IL-6 ELISA for the effect on the $IL-1{\beta}$, IL-6 secretion of human gingival fibroblast. Also gelatinase zymography and gelatin dissolubility test for the activation of MMP-2 secreted by gingival fibroblast. The results were as follows. 1. The effect of TDC and TLC on IL-6 secretion of human gingival fibroblast showed statistically significant increase of IL-6 secretion in the TDC and TLC treated group compared to no treatment group(p<0.05) . 2. The amount of $IL-1{\beta}$ secretion was below the lower limit and there was no difference in the $IL-1{\beta}$ secretion of gingival fibroblast between TDC, TLC treated group and no treatment group. 3. The active form of pro MMP-2 with 72 kDa molecular weight was activated in both TDC and TLC treated group and clear band was appeared at 62kDa site on the zymography. 4. Gelatin dissolubility of MMP-2 secreted by gingival fibroblast was higher in TDC and TLC treated group compared to no treatment group(p<0.05). 5. In the TDC treated group, serine protease of T. denticola affect gelatin dissolubility. But in the TLC treated group gelatin was degraded by only MMP secreted by gingival fibroblast. Regarding to the above results, TDC and TLC have an effect on the IL-6 secretion increase of human gingival fibroblast and appears to activate pro MMP-2 which degrades collagen.